Using a bioaerosol personal sampler in combination with real-time PCR analysis for rapid detection of airborne viruses
Using a bioaerosol personal sampler in combination with real-time PCR analysis for rapid detection of airborne viruses
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DOI：
10.1111/j.1462-2920.2006.01226.x
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发表时间：
2007-04-01
影响因子：
5.1
通讯作者：
Khromykh, Alexander A.
中科院分区：
文献类型：
--
作者：
Pyankov, Oleg V.;Agranovski, Igor E.;Khromykh, Alexander A.
我们最近开发了一种新的个人采样器，并证明了其检测空气中可存活的微生物包括细菌、真菌和病毒的可行性。为了加快耗时的分析程序，包括2-5天的生物测试，我们使用了实时PCR协议与个人采样器相结合来收集空气传播的病毒。这种方法的优点是，如果通过聚合酶链式反应检测到空气中存在特定病原体，则可以用更耗时的生物学方法进一步分析剩余的采集液，以估计空气中感染/活微生物的数量。由于自然环境中生物气溶胶的采样可能与环境空气中普遍存在的一系列微生物的大量污染有关，因此需要通过靶向聚合酶链式反应分析对检测的特异性进行调查。本文介绍了用实时荧光定量聚合酶链式反应技术检测高浓度细菌和真菌污染的环境空气中流感病毒的研究结果。联合采样-聚合酶链式反应检测方法对空气中其他微生物浓度升高的不稳定空气传播病毒的快速(类似于2.5h)和高特异性(无交叉反应)的鉴定是完全可行的。
We have recently developed a new personal sampler and demonstrated its feasibility for detection of viable airborne microorganisms including bacteria, fungi and viruses. To accelerate the time-consuming analytical procedure involving 2-5 days of biological testing, we employed a real-time PCR protocol in conjunction with the personal sampler for collection of airborne viruses. The advantage of this approach is that if the presence of a particular pathogen in the air is detected by the PCR, the remaining collecting liquid can be further analysed by more time-consuming biological methods to estimate the number of airborne infectious/live microorganisms. As sampling of bioaerosols in natural environments is likely to be associated with substantial contamination by a range of microorganisms commonly existing in an ambient air, an investigation of the specificity of detection by targeted PCR analysis is required. Here we present the results of the study on the detection of Influenza virus in the ambient air contaminated with high concentrations of bacteria and fungi using real-time PCR protocol. The combined sampling PCR detection method was found to be fully feasible for the rapid (similar to 2.5 h) and highly specific (no cross-reactivity) identification of the labile airborne virus in the air containing elevated concentrations of other microorganisms.